| RRC ID |
81945
|
| Author |
Inagaki M, Abe N, Li Z, Nakashima Y, Acharyya S, Ogawa K, Kawaguchi D, Hiraoka H, Banno A, Meng Z, Tada M, Ishida T, Lyu P, Kokubo K, Murase H, Hashiya F, Kimura Y, Uchida S, Abe H.
|
| Title |
Cap analogs with a hydrophobic photocleavable tag enable facile purification of fully capped mRNA with various cap structures.
|
| Journal |
Nat Commun
|
| Abstract |
Starting with the clinical application of two vaccines in 2020, mRNA therapeutics are currently being investigated for a variety of applications. Removing immunogenic uncapped mRNA from transcribed mRNA is critical in mRNA research and clinical applications. Commonly used capping methods provide maximum capping efficiency of around 80-90% for widely used Cap-0- and Cap-1-type mRNAs. However, uncapped and capped mRNA possesses almost identical physicochemical properties, posing challenges to their physical separation. In this work, we develop hydrophobic photocaged tag-modified cap analogs, which separate capped mRNA from uncapped mRNA by reversed-phase high-performance liquid chromatography. Subsequent photo-irradiation recovers footprint-free native capped mRNA. This approach provides 100% capping efficiency even in Cap-2-type mRNA with versatility applicable to 650 nt and 4,247 nt mRNA. We find that the Cap-2-type mRNA shows up to 3- to 4-fold higher translation activity in cultured cells and animals than the Cap-1-type mRNA prepared by the standard capping method.
|
| Volume |
14(1)
|
| Pages |
2657
|
| Published |
2023-5-11
|
| DOI |
10.1038/s41467-023-38244-8
|
| PII |
10.1038/s41467-023-38244-8
|
| PMID |
37169757
|
| PMC |
PMC10175277
|
| MeSH |
Animals
Cells, Cultured
Protein Biosynthesis*
RNA Caps* / metabolism
RNA, Messenger / genetics
RNA, Messenger / metabolism
|
| IF |
12.121
|
| Resource |
| Human and Animal Cells |
HeLa.S3(RCB1525)
HeLa(RCB0007) |
